This project will use phenanthrene and phenanthrene-9,10-oxide (PhO) as model compounds to assess the value of HPLC separation with ELISA detection in studying the metabolism of and DNA adduct formation by polycyclic aromatic hydrocarbons. A library of monoclonal antibodies will be examined for chiral specificity. Since two optical isomers can have profoundly different biological effects, there is a substantial need for chirally specific analytical methodologies including ELISAs. HPLC methods will be developed to separate the urinary metabolites with highly sensitive detection being done by ELISA. Detection limits and the ability to make antibody cross reactivity an advantage by detecting multiple species will be assessed. PhO will be reacted with DNA nucleosides and DNA, with the latter being cleaved to nucleosides and adducts. Methods will be developed to separate individual species by HPLC and detect them with ELISAs. Chiral interactions will be of particular interest. The goal will be to demonstrate methods useful in the long term in the simple, rapid, non-invasive, and extremely sensitive detection and quantitation of human exposure to environmental mutagens and carcinogens (molecular epidemiology).